Metatranscriptomics reveals overall active bacterial composition in caries lesions

Journal of Oral Microbiology

Alex Mira²  Áurea Simón-Soro¹  Miriam Guillén-Navarro¹ 

[1] FISABIO Foundation, Center for Advanced Research in Public Health;FISABIO Foundation, Center for Advanced Research in Public Health

关键词: 16S rRNA;    pyrosequencing;    Streptoccus mutans;    polymicrobial disease;    tissue-dependent hypothesis;    caries etiology;   

DOI  :  

来源: Co-Action Publishing

【 摘 要 】

Background:Identifyingthemicrobialspeciesincarieslesionsisinstrumentaltodeterminetheetiologyofdentalcaries.However,asignificantproportionofbacteriaincariouslesionshavenotbeencultured,andtheuseofmolecularmethodshasbeenlimitedtoDNA-basedapproaches,whichdetectbothactiveandinactiveordeadmicroorganisms.Objective:ToidentifytheRNA-based,metabolicallyactivebacterialcompositionofcarieslesionsatdifferentstagesofdiseaseprogressioninordertoprovidealistofpotentialetiologicalagentsoftoothdecay.Design:Non-cavitatedenamelcarieslesions(n=15)anddentincarieslesionssamples(n=12)werecollectedfrom13individuals.RNAwasextractedandcDNAwasconstructed,whichwasusedtoamplifythe16SrRNAgene.Theresulting780bppolymerasechainreactionproductswerepyrosequencedusingTitanium-pluschemistry,andthesequencesobtainedwereusedtodeterminethebacterialcomposition.Results:Ameanof4,900sequencesofthe16SrRNAgenewithanaveragereadlengthof661bpwasobtainedpersample,givingacomprehensiveviewoftheactivebacterialcommunitiesincarieslesions.Estimatesofbacterialdiversityindicatethatthemicrobiotaofcavitiesishighlycomplex,eachsamplecontainingbetween70and400metabolicallyactivespecies.Thecompositionofthesebacterialconsortiavariedamongindividualsandbetweencarieslesionsofthesameindividuals.Inaddition,enamelanddentinlesionshadadifferentbacterialmakeup.Lactobacilliwerefoundalmostexclusivelyindentincavities.Streptococciaccountedfor40%ofthetotalactivecommunityinenamelcaries,and20%indentincaries.However,Streptococcusmutansrepresentedonly0.02–0.73%ofthetotalbacterialcommunity.Conclusions:Thedataindicatethattheetiologyofdentalcariesistissuedependentandthatthediseasehasaclearpolymicrobialorigin.Thelowproportionofmutansstreptococcidetectedconfirmsthattheyareaminorityandquestionsitsimportanceasthemainetiologicalagentoftoothdecay.Futureexperimentalworkshouldbeperformedtoconfirmthecariogenicityoftheidentifiedbacteria.Keywords:16SrRNA;pyrosequencing;Streptococcusmutans;polymicrobialdisease;tissue-dependenthypothesis;cariesetiology(Published:24October2014)Citation:JournalofOralMicrobiology2014,6:25443-http://dx.doi.org/10.3402/jom.v6.25443

【 授权许可】

Unknown